NHDL, a recombinant VL/VH hybrid antibody control for IgG2/4 antibodies
Corinna Lau,
Martin Berner McAdam,
Grethe Bergseth,
Algirdas Grevys,
Jack Ansgar Bruun,
Judith Krey Ludviksen,
Hilde Fure,
Terje Espevik,
Anders Moen,
Jan Terje Andersen,
Tom Eirik Mollnes
Affiliations
Corinna Lau
Research Laboratory, Nordland Hospital Trust, Bodø, Norway
Martin Berner McAdam
Department of Immunology, Oslo University Hospital-Rikshospitalet, and Centre for Immune Regulation, Oslo, Norway
Grethe Bergseth
Research Laboratory, Nordland Hospital Trust, Bodø, Norway
Algirdas Grevys
Department of Immunology, Oslo University Hospital-Rikshospitalet, and Centre for Immune Regulation, Oslo, Norway
Jack Ansgar Bruun
Department of Medical Biology, Proteomics Platform, University of Tromsø, Tromsø, Norway
Judith Krey Ludviksen
Research Laboratory, Nordland Hospital Trust, Bodø, Norway
Hilde Fure
Research Laboratory, Nordland Hospital Trust, Bodø, Norway
Terje Espevik
Centre of Molecular Inflammation Research, and Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway
Anders Moen
Department of Biosciences, Proteomics core facility, University of Oslo, Oslo, Norway
Jan Terje Andersen
Department of Immunology, Oslo University Hospital-Rikshospitalet, and Centre for Immune Regulation, Oslo, Norway
Tom Eirik Mollnes
Research Laboratory, Nordland Hospital Trust, Bodø, Norway
The mechanism of action of recombinant IgG2/4 antibodies involves blocking of their target without the induction of effector functions. Examples are eculizumab (Soliris®), which is used clinically to block complement factor C5, as well as anti-human CD14 (r18D11) and anti-porcine CD14 (rMIL2) produced in our laboratory. So far, no proper IgG2/4 control antibody has been available for controlled validation of IgG2/4 antibody functions. Here, we describe the design of a recombinant control antibody (NHDL), which was generated by combining the variable light (VL) and heavy (VH) chains from two unrelated specificities. NHDL was readily expressed and purified as a stable IgG2/4 antibody, and showed no detectable specificity toward any putative antigen present in human or porcine blood. The approach of artificial VL/VH combination may be adopted for the design of other recombinant control antibodies.
